Low/negative expression of DDX3 might predict poor prognosis in non-smoker patients with oral cancer.

OBJECTIVE: DDX3 has diverse biological functions in translation control, cell growth regulation, and tumor progression. Oral squamous cell carcinoma (OSCC) is a common malignant tumor worldwide with a poor clinical prognosis. The impact of DDX3 expression in OSCC is seldom discussed. MATERIALS AND METHODS: Tumor tissues and adjacent normal tissues were obtained from 324 patients with OSCC. In this study, we used immunohistochemical staining methods to investigate the associations between DDX3 expression and the clinicopathological characteristics of OSCC. RESULTS: Low/negative DDX3 expression in tumor cells was significantly associated OSCC patient characteristics including male gender (P < 0.001), smoking (P < 0.001), alcohol consumption (P < 0.001), betel quid chewing (P = 0.002), poor relapse-free survival (P = 0.001), and poor overall survival (OS) (P = 0.001). Patients with low/negative DDX3 expression, and particularly non-smoker OSCC patients, had significantly worse OS as defined by the log-rank test (P = 0.020 for all cases; P = 0.008 for non-smoker patients). In non-smoker patients with OSCC, low/negative DDX3 expression in tumor cells was associated with poor prognosis (P = 0.024) and a 3.802-fold higher death risk, as determined by Cox regression. CONCLUSIONS: Low/negative DDX3 expression in tumor cells was significantly associated with aggressive clinical manifestations and might be an independent survival predictor, particularly in non-smoker patients with OSCC.

The prognostic significance of APC gene mutation and miR-21 expression in advanced-stage colorectal cancer.

AIM: Colorectal cancer (CRC) is the second commonest cause of cancer death in Taiwan. Although numerous genes have been associated with tumorigenesis in colorectal cancer, only a few have been validated and used as biomarkers for predicting clinical outcome. The aim of this study was to analyse the association of APC gene mutation and miR-21 expression with clinical outcome in CRC patients. METHOD: In total, 195 colorectal cancer patients were enrolled in a single medical centre between 2003 and 2007. APC gene mutation and expression of APC and miR-21 were analysed by direct DNA sequencing and real-time reverse transcription polymerase chain reaction. The primary outcome included 5-year overall survival and univariate (Kaplan-Meier) and multivariate (Cox regression) analyses of prognostic factors. RESULTS: The results showed that 66 (33.8%) of 195 tumour tissues contained an APC mutation. The predominant APC gene variations were deletion mutations (50.0%). APC gene expression was low in CRC and negatively correlated with miR-21 expression and gene mutation. In advanced-stage cancer, patients with APC mutation/high miR-21 had poorer overall survival rates than those with APC mutation/low miR-21, APC wild-type/high miR-21 and APC wild-type/low miR-21. CONCLUSION: In Taiwan, downregulation of the APC gene in CRC correlated with gene mutation and miR-21 upregulation. APC mutation and miR-21 expression could be used to predict the clinical outcome of CRC, especially in patients with advanced disease.

Increased expression of PRL-1 protein correlates with shortened patient survival in human hepatocellular carcinoma.

BACKGROUND: The purposes of the current study were to investigate whether overexpression of the PRL-1 is clinically relevant to hepatocellular carcinoma (HCC) and whether expression patterns of PRL-1 in HCC have diagnostic and prognostic value. METHODS: Immunohistochemistry analysis was performed for PRL-1 in 60 HCC samples. The data were correlated with clinicopathological features. The univariate and multivariate survival analyses were also performed to determine their prognostic significance. RESULTS: PRL-1 protein was overexpressed (83%) in HCC as compared with the adjacent normal tissue. PRL-1 expression was not influenced by chronic alcohol exposure or cirrhosis. High expression of PRL-1 was correlated with smoking (p=0.012), cirrhosis (p=0.047) and histological grade (p=0.055). The Kaplan-Meier survival curves showed that high PRL-1 expression related to a poor survival with statistical significance (I vs. III, p=0.010; II vs. III, p=0.001). Univariate analysis showed that PRL-1 expression was associated with tumour size, stage and PRL-1 score. Multivariate analysis revealed that the PRL-1 protein expression level was an independent factor for overall survival (HR, 5.367; 95% CI, 2.270-12.692; p=0.001). This is the first demonstration that the expression level of PRL-1 is correlated with tumour progression and prognosis in HCC. CONCLUSIONS: Along with other results, the PRL-1 protein is a candidate biomarker and a potential target for novel therapies against human HCC progression.

Experimental treatment of bilateral fetal chylothorax using in-utero pleurodesis.

OBJECTIVE: To assess the use and efficacy of in-utero pleurodesis for experimental treatment of bilateral fetal chylothorax. METHODS: This was a study of 78 fetuses with bilateral pleural effusion referred to three tertiary referral centers in Taiwan between 2005 and 2009. Fetuses were karyotyped following amniocentesis and the lymphocyte ratio in the pleural effusion was determined following thoracocentesis. Forty-nine (62.8%) fetuses had a normal karyotype and were recognized to have fetal chylothorax; of these, 45 underwent intrapleural injection of 0.1KE OK-432 per side per treatment. We evaluated clinical (hydrops vs. no hydrops) and genetic (mutations in the reported lymphedema-associated loci: VEGFR3, PTPN11, FOXC2, ITGA9) parameters, as well as treatment outcome. Long-term survival was defined as survival to 1 year of age. RESULTS: The overall long-term survival rate (LSR) was 35.6% (16/45); the LSR for non-hydropic fetuses was 66.7% (12/18) and for hydropic fetuses it was 14.8% (4/27). If we included only fetuses with onset of the condition in the second trimester, excluding those with onset in the third trimester, the LSR decreased to 29.4% (10/34). Notably, 29.6% (8/27) of hydropic fetuses had mutations in three of the four loci examined. CONCLUSIONS: OK-432 pleurodesis appeared to be an experimental alternative to the gold-standard technique of thoracoamniotic shunting in non-hydropic fetal chylothorax. In hydropic fetuses, pleurodesis appeared less effective.

Distribution of lysosome-associated membrane proteins-1 and -2, and cathepsin D in eosinophilic granular bodies: possible relationship to cyst development in pilocytic astrocytomas.

Pilocytic astrocytomas are usually cystic; cyst formation within these tumours may result in increased intracranial pressure, due to the effect of their mass, and contribute to cerebral damage. Eosinophilic granular bodies (EGBs) are produced abundantly in pilocytic astrocytomas but their role in disease progression remains unknown. Immunohistochemistry studies showed EGBs to exhibit pronounced reactivity to antibodies against lysosome-associated membrane proteins (LAMP)-1 and LAMP-2, and the lysosomal enzyme cathepsin D. Both LAMP-1 and LAMP-2 showed peripheral rim and granular staining patterns. The EGBs were scattered widely across cysts and, where EGBs aggregated in clusters, were usually close to areas of fluid in the cysts. Most EGBs had nuclei either attached or close by, indicating that the EGBs may be derived from anucleated astrocytes. The results suggest that EGBs, together with other factors, may play a role in the development of cysts in pilocytic astrocytomas.

Increased serum osteopontin is a risk factor for osteoporosis in menopausal women.

SUMMARY: Osteopontin (OPN)-deficient mice are resistant to ovariectomy-induced osteoporosis. Therefore, we hypothesized that women with OPN overexpression may show less resistance to postmenopausal osteoporosis. In this study, we first demonstrated that serum OPN levels could be used as a biomarker for the early diagnosis of osteoporosis in postmenopausal women. INTRODUCTION: Animal studies indicate that OPN-deficient mice are resistant to ovariectomy-induced osteoporosis. METHODS: From 2004 to 2006, 124 women over the age of 45 were enrolled in a menopausal group, while another 95 women, from 25 to 45 years of age with regular menstruation, were enrolled into a childbearing age group. The serum concentrations of OPN were calculated using the enzyme-link immunosorbent assay method, and bone mineral densities were determined with dual energy X-ray absorptiometry. RESULTS: Serum OPN levels had a significant positive correlation with age (menopausal group, p < 0.0001) and a negative correlation with body weight, height, hip bone mineral density, and T-scores in the menopausal group. In contrast, there was a positive correlation with the E2 concentration and height, but there was no significant association with the above variables in the childbearing age group. Additionally, high serum OPN levels (>14.7 ng/ml) was a significant risk factor causing menopausal osteoporosis (odds ratio = 2.96, 95% confidence interval, 1.055-8.345). CONCLUSION: Serum OPN levels could be used as a biomarker for the early diagnosis of osteoporosis in postmenopausal women.

Genetic evaluation and management of fetal chylothorax: review and insights from a case of Noonan syndrome.

Fetal chylothorax is one of a very few syndromes that can be treated in utero with thoracoamniotic shunting or pleurodesis by OK-432 as two major therapeutic modalities. We report on a fetus with Noonan syndrome and a missense mutation c.182A > C (p.Asp61Ala) of PTPN11 who responded poorly to antenatal pleurodesis by OK-432. Based on our previous publication and this case study, we propose that fetal chylothorax of a distinct genetic origin may respond poorly to OK-432 pleurodesis.

Cortactin overexpression in the esophageal squamous cell carcinoma and its involvement in the carcinogenesis.

The aim of this study is to examine whether dysregulated expression of cortactin occurs in esophageal squamous cell carcinoma (ESCC) and is involved in the development of ESCCs. An immunohistochemistry study for cortactin expression was performed on 46 pairs of surgically resected non-tumor and ESCC tumor tissues and murine tumors of esophagi induced by a carcinogen. The results show increased cortactin expression in 20 and in 22 to a lesser extent, out of a total 46 ESCC tumor tissues. Increased cortactin was also detected in the premalignant lesions, the early stage dysplasia and carcinoma in situ, of ESCC tumor tissues. Differential polymerase chain reaction results showed slight increases in the EMS1 gene only in two of 10 ESCC tumor tissues, suggesting that EMS1 gene amplification is not the only mechanism for cortactin overexpression. In the mouse model induced by treatment with 4-nitroquinoline 1-oxide and arecoline, increased cortactin was detected in the epithelia with hyperkeratosis, papillomas, and ESCCs with invasion into the submucosa, respectively. Overall, we observed cortactin overexpression in early and late stages of human ESCCs and carcinogen-induced murine ESCCs, suggesting a role for cortactin in esophageal carcinogenesis.

Novel transmembrane GTPase of non-small cell lung cancer identified by mRNA differential display.

The technique of differential display was used previously to profile the gene expression patterns of non-small cell lung cancer, and several genes differentially expressed were thus identified. In this report, we demonstrate that a DNA fragment of 347-bp length, up-regulated in tumor tissues, showed 100% sequence similarity to human cDNA FLJ20693 for a 370-residue protein. The gene product of cDNA FLJ20693 was postulated to be a shorter isoform of transmembrane GTPase, termed TG370, based upon the results of searching for sequence homology. The nucleotide sequence alignment also indicated that the cDNA FLJ20693 and the cDNA for 741-residue human mitofusin 1 (TG741) possibly resulted from the event of alternative splicing from which a 127-bp region was retained in the latter. Analysis of the genome sequence confirmed the speculation that both cDNAs were mapped to the same chromosomal position composing of 18 exons, of which the 127-bp region of TG741 constituted exon 11. The alternative splicing in all lung cancer cell lines was also observed to occur nearly in all tissue specimens examined. The up-regulated expression of transmembrane GTPase was subsequently found in tumor tissues from at least five of seven non-small cell lung cancer patients. Also, a distinct PCR product was initially detected in cell line H520, and further sequence analysis identified the presence of the 86-bp region mapped to the genome sequence immediately followed by exon 10. To evaluate the retention of 86-bp region, it was found that, besides the predicted 486-bp product, an unexpected 332-bp product was concomitantly observed and identified as the result of exon 8 deletion. The expression and subcellular localization of the full-length TG741 and other shorter isoforms were detected by flow cytometry using three polyclonal antibodies. It was concluded that the full-length TG741 located at plasma membrane with its NH(2)-terminal domain exposed extracellularly and the shorter isoforms retained at cytosol. Finally, the up-regulation of transmembrane GTPase in tumor tissues was further illustrated using immunohistochemical staining.

Thyroid metastasis from intraductal papillary-mucinous carcinoma of the pancreas. A case report.

BACKGROUND: Intraductal papillary-mucinous carcinoma (IPMC) of the pancreas is a newly identified clinicopathologic entity of the exocrine pancreas. It has been considered a slowly growing and less-aggressive carcinoma with a favorable prognosis. There have been only a few documents reporting its distant metastasis and cytologic features, with no report of thyroid metastasis until the present. CASE: A case of IPMC occurred in a 45-year-old male, who was admitted with rapid growth and tenderness of the thyroid. Abdominal computed tomography showed the typical cystic dilatation of IPMC with adjacent organ metastasis. Fine needle aspiration of the thyroid yielded papillary fronds of carcinoma cells with nuclear pleomorphism, abundant cytoplasm and prominent nucleoli in a mucinous background. Immunohistochemical findings from the skin and thyroid characterized the papillary-mucinous carcinoma as having originated in the pancreas. CONCLUSION: This case suggests that papillary carcinoma fronds aspirated from the thyroid should be further differentiated from the primary site and that a pleomorphic nucleus in a mucinous background is a useful feature to exclude a thyroid origin. Before this, distant metastasis of IPMC to the skin and thyroid has not been reported. The prognosis of IPMC with wide, distant metastasis at an advanced stage is poor.

Mutation analysis of the Bcl 10 gene in hepatocellular carcinoma.

The Bcl 10 gene was recently discovered to be involved in the pathogenesis of lymphoma of mucosa-associated lymphoid tissue and several types of tumorous cell lines. We examined the mutation of Bcl 10 gene in 31 hepatocellular carcinomas along with their corresponding non-tumorous tissues by single strand conformation polymorphism (SSCP) and direct sequencing. The results showed that 11.3% chromosomes had codon 5 GCA to TCA mutation, 4.8% chromosomes had codon 8 CTC to CTG mutation, and 12.9% chromosomes had codon 213 GGA to GAA mutation. These mutations were found not only in the hepatoma tissues but also in paired non-cancerous tissues and the normal population. We suggest that these three changes are polymorphisms, and there is no relationship with the development of hepatocellular carcinoma.

No evidence of correlation between mutation at codon 531 of src and the risk of colon cancer in Chinese.

The protein tyrosine kinase activity of c-src proto-oncogene product, pp60(c-src), is elevated in a number of human cancers, including colon cancer. Phosphorylation of human pp60(c-src) carboxy-terminal tyrosine 530 suppresses its kinase activity. A recent report suggested that the risk of colon cancer is higher for those who carry a C-->T transition mutation on codon 531 (Gln-531-->Amber-531) of src gene. This mutation caused a prematured translation termination and up-regulated the kinase activity. To examine whether this mutation could be a risk factor for colon carcinoma in the Chinese population, we used the same PCR-based assay to analyze src genotypes of 131 colon cancers and other various types of carcinoma. No mutation was detected in all specimens that were screened in this study. Thus, mutation at Gln-531 of src gene does not seem to be involved in the development of colon cancer in Chinese ethnicity.

Mutation analysis of the putative tumor suppressor gene PTEN/MMAC1 in hepatocellular carcinoma.

Loss of heterozygosity of chromosome 10q has been reported in hepatoma. Areas with a high rate of loss of genetic material could harbor putative tumor suppressor genes. PTEN/MMAC1, a candidate tumor suppressor gene located at chromosome 10q23.3, has recently been identified and found to be homozygously deleted or mutated in several different types of human tumors. To determine whether the PTEN/MMAC1 gene is a target of 10q loss of heterozygosity in hepatoma, we examined 42 primary hepatomas for mutations in PTEN/MMAC1 by using nested reverse transcriptase polymerase chain reaction (RT-PCR) of the RNA and single-stranded conformation polymorphism (SSCP) analysis of all genomic exons. Although 2 of 42 hepatoma tissues had aberrant transcripts, 5 matched noncancerous liver tissues also had aberrant transcripts. Southern blot analysis of the entire genomic DNA revealed no genomic change. Therefore, like the TSG101 or FHIT gene, aberrant transcripts of PTEN/MMAC1 using the nested RT-PCR method were a common phenomenon for both cancerous and noncancerous liver tissues, which may not be related to oncogenesis. None of the 42 cases had small deletions, point mutations, or insertions. Our results suggest that the PTEN/MMAC1 gene may not play a role in the pathogenesis of hepatoma.

Do the CK18 related proteins change in general in epithelial cancers?

The modulation of cytokeratin 18 during tumor transformation in hepatoma had been previously recognized through a series of biochemical and immunological approaches. Expression of cytokeratin 18 in transitional cell carcinoma comparing with hepatoma was investigated using the hepatoma transformation model. CK18 related molecules were found. In the present study, we design various epithelial cancers with the same model. CK18 related molecules were all evident. Therefore, we suggest that CK18 related proteins would play an important role in tumorigenesis of epithelial cancers.

Subcutaneous sparganosis--a case report and a review of human sparganosis in Taiwan.

Human sparganosis is a rare parasitic disease infected by plerocercoid larva (sparganum) of Spirometra species. It was usually diagnosed accidentally and has long been underestimated. In this report, we describe a 53-year-old woman presenting as an enlarging subcutaneous nodule in the right thigh for 3 months, which was excised in the belief that it was a lipoma. Characteristic sparganum accompanied by granulomatous inflammation, eosinophilic infiltrate and sinus tract in the subcutaneous tissue were discovered under microscopic examination of the excised tissue. Contaminated drinking water was presumed to be the infectious source. Complete excision is a curative treatment. We also review previously documented 19 human sparganosis in Taiwan to provide the clinical context for this report.

Mutation analysis of the putative tumor suppression gene PTEN/MMAC1 in sporadic breast cancer.

PTEN/MMAC1, a potential human tumor suppressor gene, has been found to have inactivating mutations in several types of cancer, including breast cancer. The incidence of breast cancer in Chinese is quite low in comparison with Caucasians, and genetic factors may play some roles. To further determine the role of PTEN/MMAC1 in breast cancer in Chinese, we used loss of heterozygosity (LOH), single strand conformation polymorphism (SSCP) with direct sequencing of variant bands, and Southern blot analysis methods to analyze mutations in PTEN/MMAC1 in 52 cases of breast cancer. None had LOH at chromosome 10q23.3. One mutation was identified, a somatic 3-base deletion, in one case. Our results suggest PTEN/MMAC1 does not play a major role in the development of sporadic breast cancer.

P53 codon 72Arg polymorphism is not a risk factor for carcinogenesis in the chinese.

The potential association of distinct polymorphism of the tumor suppressor gene p53, with an increased susceptibility to malignant transformation has been reported for various cancers. A polymorphism at codon 72 of p53 results in translation to either arginine (p53Arg) or proline (p53Pro), and recent study showed that Caucasian women with arginine form of p53 are more susceptible to HPV-associated carcinoma of the cervix. To examine whether arginine 72 could be a significant risk factor for tumor development, we used a PCR-based assay to analyze p53 genotypes of patients for several types of carcinoma. No significant difference in the frequency of p53Arg was found between normal and cancer patients, the results showed that the individuals homozygous for arginine variant were not at increased risk for cancer.